Soybean rust caused by Phakopsora pachyrhizi Syd. is a potential disease causing severe losses in yield and oil quality of soybean [Glycine max (L.) Merr.]. The physiological variability studies were carried out for 25 isolates collected from different growing environment of Karnataka, India during Kharif 2010-2012. Morphological and cultural characters of 25 isolates were studied under identical condition. Among 25 isolates, the maximum uredospore size was found in Ugarkhurd isolate (90.93  70.05 m). The differential JS-335 reacted to the maximum of eight isolates, and isolate Rayanal produced both RB and TAN reaction on most of the differentials. In the clustering of different isolates into pathotypes, the cluster analysis indicated the expression of eight isolates on differential JS-335 and there was no expression of all the isolates on differential PI-459024B. In cluster-II, nine isolates expressed on PI-200492, while one isolate expressed on JS-335. In cluster-III, 23 isolates expressed on PI-459024B and 12 isolates expressed on PI-200492. Hence, this study described the three pathotypes that causes soybean rust in India namely, Cluster I-Reddish Brown (RB) lesion producing pathotypes, Cluster II -TAN lesion producing pathotypes and Cluster III -Mixed or RB + TAN producing pathotypes. This information could further be used for identification of race pattern in Karnataka and other major rust prone areas of India.